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研究生:賴欣梅
研究生(外文):Lai Shin-Mei
論文名稱:使用高解析度DNA定序法來分析台灣九族原住民與台灣漢人HLAclassⅠA基因的遺傳多態性:應用於探討台灣原住民的起源與擴散
論文名稱(外文):Genetic Diversity in the HLA classⅠA Gene of the Nine Taiwan Aboriginal Tribes and Han Chinese as Revealed by High Resolution Sequence-Based HLA Typing: Implication for the Origins and Dispersal of the Taiwan Aborigines
指導教授:陳堯峰陳堯峰引用關係吳紹基
指導教授(外文):Chen Yao-fongWu Shao-kee
學位類別:碩士
校院名稱:慈濟大學
系所名稱:人類遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
畢業學年度:90
語文別:中文
論文頁數:80
中文關鍵詞:台灣原住民
外文關鍵詞:Taiwan aborigines
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人類白血球抗原(HLA)是目前已知的功能基因中最具多態性的基因群。因為HLA基因高度的多態性,可藉由此基因多態性的分析結果,再佐以文化、考古及語言學等證據,以了解族群間的遺傳類緣關係,並探討與重建族群的遷徙與起源。 本文使用DNA定序法(SBT),分析台灣九族原住民(阿美族、泰雅族、布農族、排灣族、卑南族、魯凱族、賽夏族、鄒族與雅美族)及台灣漢人的HLA classⅠ基因群中的HLA-A基因。等位基因頻率及單倍體(A-B及DPB1-DRB1-DRB3/4/5-A-B)頻率採用最大相似值法計算得知,再由等位基因頻率計算出遺傳距離,進而建立種系發生樹(phylogenetic tree)。 研究結果顯示,台灣原住民的等位基因數目少,等位基因頻率集中於少數的等位基因型。相對於台灣漢人而言,台灣九族原住民是同質性較高的族群,其中以雅美族的族群同質性最高。台灣漢人與台灣原住民不論在等位基因型,或等位基因頻率分佈上皆有差異。台灣原住民的HLA-A基因集中於A*2402型,以賽夏族的30.0%最低,排灣族的88.0%最高,在其他族群中平均約有50.0∼60.0%的分佈,台灣漢人則只有21.0%的分佈。另外,在泰雅和賽夏兩族中,我們發現了一個新的HLA-A等位基因型A*2420型,兩族分佈頻率分別為15.0%及13.0%。單倍體頻率與遺傳距離的分析同樣顯示出阿美族及雅美族與其他族群的遺傳關係較遠,以及台灣漢人與台灣原住民九族在HLA基因遺傳上的差異。從種系發生樹可看出台灣原住民九族可分為三大群:第一群是排灣族、布農族、鄒族及魯凱族;第二群是雅美族;第三群則是其他四族,第三群的四族似乎又可細分為三群:一是阿美族;二是泰雅族及賽夏族;三是卑南族。我們的研究中發現台灣原住民可能在不同的時間,由不同的地點來到台灣。 未來可以此研究的結果為基礎,與其他東南亞及菲律賓等地的南島語族族群,及南中國大陸的少數民族遺傳資料相比對,以釐清台灣原住民的起源與擴散問題,並期能更進一步能將此結果應用於了解整個南島語族的起源與擴散。
The polygenic HLA (human leukocyte antigen) gene complex is the most polymorphic cluster of functional genes known. The high polymorphism of HLA genes makes these functional genes ideal genetic markers for studying inter-relationships among various human populations. Combined the results of HLA polymorphism with evidences from socio-cultural, archaeological and linguistic studies, we may understand better the origin and dispersal of human populations. In this study we have used sequence-based typing (SBT) to analyze the degree of polymorphism of classΙ HLA-A gene in the nine Taiwan aboriginal tribes (Amis, Atayal, Bunun, Paiwan, Puyuma, Rukai, Saisiat, Tsou, and Yami) and Taiwan Han Chinese. The allele frequencies of each population are estimated by a maximum-likelihood method and used to calculate genetic distances between pairwise populations according to Nei’s formula. Phylogenetic tree is then constructed from these genetic distances. The results show limited genetic diversity of Taiwan aborigines, especially the Yami tribe. The distributions of allele frequencies of Taiwan aborigines versus Han Chinese are different. All the Taiwan aboriginal tribes have a high percentage of A*2402 allele ranging from 30% in Saisiat to 88% in Paiwan and Taiwan Han Chinese has 21%. We found a new HLA-A allele A*2420 in Atayal and Saisiat, and the allele frequencies are 15% and 13%, respectively. Analysis of allele frequencies and genetic distances of HLA-A gene reveals that Amis and Yami are different from other Taiwan aboriginal tribes. From the phylogenetic tree, nine Taiwan aboriginal tribes are divided into three clusters:the first one includes Paiwan, Bunun, Tsou and Rukai; the second one is Yami; and the third one includes the other four tribes. The third cluster can be further divided into three groups: the first one is Amis; the second includes Atayal and Saisiat; and the third includes Puyuma. In the future, when there are more genetic data from the Philippine and Southeast Asia populations, we may understand much more about the origin and dispersal of Taiwan aborigines in particular, and the Austronesians in general.
總 目 次 中文摘要………………………………………………………………Ⅰ 英文摘要………………………………………………………………Ⅱ 圖目次…………………………………………………………………Ⅲ 表目次…………………………………………………………………Ⅳ 第一章 導論…………………………………………………………1 1.1 台灣原住民介紹…………………………………………1 1.1.1 族群的分佈…………………………………………1 1.1.2 語言特徵……………………………………………2 1.1.3 社會組織與文化…………………………………3 1.1.4 族群起源……………………………………………5 1.2 人類白血球抗原 (Human Leukocyte Antigen)……………8 1.2.1 簡介…………………………………………………8 1.2.2 分子結構……………………………………………8 1.2.3 功能…………………………………………………9 1.2.4 HLA與疾病的關係………………………………10 1.2.5 HLA與移植………………………………………...11 1.2.6 HLA等位基因之命名……………………………...11 1.2.7 HLA基因分型方法………………………………...12 1.2.8 HLA基因多態性在遺傳人類學的應用…………...14 1.3 台灣原住民遺傳標記研究的回顧…………………………15 1.4 研究目的……………………………………………………19 第二章 實驗材料與方法…………………………………………….20 2.1 研究對象……………………………………………………20 2.2 實驗方法……………………………………………………21 2.2.1 DNA萃取及定量…………………………………...21 2.2.2 HLA-A基因分型…………………………………...21 2.2.3 基因序列分析與比對……………………………….21 2.2.4 資料分析…………………………………………….22 2.2.5 TA cloning…………………………………………...22 第三章 實驗結果…………………………………………………….23 3.1 HLA-A基因………………………………………………....23 3.2 新HLA-A等位基因型A*2420…………………………….25 第四章 討論………………………………………………………….26 References……………………………………………………………....32 Appendix A……………………………………………………………..67 Appendix B……………………………………………………………..74 Appendix C……………………………………………………………..75 Appendix D……………………………………………………………..76 Appendix E……………………………………………………………...77 Appendix F……………………………………………………………...78 Appendix G……………………………………………………………..79 Appendix H……………………………………………………………..80
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